JP2003160481A - N-propargyl-1-amino indane, its salt, composition and use thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain R(+)-N-propargyl-1-aminoindane, its pharmaceutically acceptable salt and a pharmaceutical composition containing the same. <P>SOLUTION: This method for treating patients suffering from Parkinson's disease, a memory disorder, dementia, depression, hyperactive syndrome, an affective illness, a neurodegenerative disease, a neurotoxic injury, brain ischemia, a head trauma injury, a spinal trauma injury, schizophrenia, an attention deficit disorder, multiple sclerosis or withdrawal symptoms comprises using R(+)-N- propargyl-1-aminoindane or its pharmaceutically acceptable salt. This method for preventing nervous damage of patient is provided. Finally this method for producing R(+)-N-propargyl-1-aminoindane, its salt and racemic isomer is provided. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

This application is a continuation of US application no. 08 / 139,517, filed October 18, 1993, the contents of which are incorporated herein by reference.

[0002] The background of the present invention I. The present invention is in the field of selective irreversible inhibitors of monoamine oxidase enzyme (hereinafter MAO), which is the selective irreversible form of the B-form of monoamine oxidase enzyme (hereinafter MAO-B). The R (+) enantiomer of the inhibitor N-propargyl-1-aminoindan (also referred to herein as PAI).
I will provide a. The present invention also relates to a pharmaceutical composition containing [R] (+)-PAI, in particular Parkinson's disease,
Memory disorder, dementia, depression, hyperactivity syndrome, emotional illness,
Neurodegenerative disease, neurotoxic injury, cerebral ischemia, traumatic head injury, traumatic spinal injury, schizophrenia, attention deficit disorder,
Also provided are those that are effective in treating multiple sclerosis and withdrawal symptoms.

II. Parkinsonism is widely believed to result from the degeneration of presynaptic dopaminergic neurons in the brain, followed by a decrease in the amount of the released neurotransmitter dopamine. Thus, insufficient dopamine release leads to the onset of spontaneous muscle control injury, which is a sign of Parkinsonism.

Various methods of treating Parkinson's disease have been established and are currently widely used, including, for example, the administration of L-dopa with a decarboxylase inhibitor such as L-carbidopa or benserazide. ing. Decarboxylase inhibitors protect the L-dopa molecule from peripheral decarboxylation, thereby ensuring uptake of L-dopa by the remaining dopaminergic neurons in the striatum of the brain. Where L
-Dopa is converted to dopamine, increasing the level of dopamine in these neurons. Thus, in response to physiological stimuli, these neurons can release large amounts of dopamine at levels close to those normally required. Thus, treatment with L-dopa alleviates the symptoms of the disease and contributes to the well-being of the patient.

However, L-dopa treatment has drawbacks, the main one being that its effect is optimal only during the first few years of treatment. After this period, the clinical response decreases, movement abnormalities, and fluctuations in drug efficacy throughout the day (“on-
Off effect ”) and adverse side effects including psychiatric symptoms such as confusion, paranoia and hallucinations. L-
This reduction in dopa treatment is due to spontaneous progression of the disease, increased dopamine production or altered dopamine receptors as a result of increased levels of dopamine metabolites, and L
-Due to many factors, including the pharmacokinetic problem of dopa absorption (Review, Youdim et al., Progress in Medicinal
Chemistry, 21 , 138-167 (1984)).

Various treatments have been devised to overcome the disadvantages of treatment with L-dopa, in which L-dopa reduces newly formed metabolites of dopamine. Combined with MAO inhibitors for the purpose (eg Chiese, published March 2, 1989,
P., U.S. Pat. No. 4,826,875).

MAO exists in two forms known as MAO-A and MAO-B, which are selective for different substrates and inhibitors. For example, MAO
-B metabolizes substrates such as 2-phenylethylamine more effectively and is selectively and irreversibly inhibited by (-)-deprenyl as shown below.

However, L-DOPA and MAO-A and MA
Treating in combination with both inhibitors of OB
As these treatments lead to adverse side effects associated with increased levels of catecholamines throughout the brainstem,
It should be noted that it is not desired. Moreover, complete inhibition of MAO also means that MAO acts by sympathomimetic amines such as tyramine (this is the so-called "cheese effect").
Lead to. ), Which is not desirable (review, Youdim e
t al., Handbook of Experimental Pharmacology, ed.
by Trendelenburg and Weiner, Springer-Verlag, 90,
ch. 3 (1988)). Since MAO-B has been shown to be the predominant form of brain MAO, inhibitors that are selective for this form will reduce dopamine degradation on the one hand and systemic inhibition of total MAO on the other hand. Considered to be a possible means to minimize sexual effects.

Many inhibitors of MAO are chiral molecules. One enantiomer often exhibits some stereoselectivity in its relative effectiveness at MAO-A and -B, but the configuration of a given enantiomer is MAO-A.
And MAO-B are less selective than their enantiomers.

Table I shows the IC ₅₀ (mmol) of enantiomeric pairs of propargylamine in the production of MAO in rat brain.
/ L) are listed. These results showed a slight difference in the effectiveness of MAO-B inhibition between the R and S enantiomers (B. Hazelhoff, et al., Naunyn-Schmeideberg's Arc.
h. Pharmacol., 330 , 50 (1985)). Both enantiomers are selective for MAO-B. In 1967, Magya
R, et al., in inhibiting oxidative deamination of tyramine by rat brain homogenates, R-
It has been reported that (-)-deprenyl is 500 times more potent than the S-(+) enantiomer (K. Magyar, et a.
l., Act. Physiol. Acad. Sci., Hung., 32, 377 (196
7)).

In rat liver homogenates, R-deprenyl was only about 15 times stronger than the S enantiomer. In other pharmacological activity assays, such as for inhibition of tyramine uptake, deprenyl shows different stereoselectivities. Form S is a more potent epimer in some cases (J. Knoll and K. Magyar, Advances
in Biochemical Psychopharmacology, 5 , 393 (197
2)).

N-methyl-N-propargyl-1-aminotetralin (2-MPAT) is a structurally close analog of deprenyl. The absolute stereochemistry of 2-MPAT has not been determined. However, the (+)-isomer is MAO-B
And the (-) isomer is selective for MAO-A. The difference in potency between the enantiomers of 2-MPAT is less than 5-fold (B. Hazelhoff, et a
l., id .). The ethane thiomers of N-propargyl-1-aminotetralin (1-PAT) are similar in activity. The lack of data in Table I indicating a clear structural activity relationship between the isolated (+)-or (−)-2-MPATs makes it possible to predict their absolute stereochemistry. It has become difficult.

After extensive computer modeling, Poly
merpoulos is (R) -N-methyl-N-propargyl-1-aminoindan (R-1-MPAI)
It was recently predicted to be more potent than (S) as a -B inhibitor (E. Polymerpoulos, Inhibitors of Monoamin
e Oxidase B, I. Szelenyi, ed., Birkhauser Verlag,
p. 110 (1993)). However, the experiments shown show that R-1-M
MAO-B with PAI slightly stronger than S-1-MPAI
, But it is a more potent inhibitor of MAO-A. Both the selectivity between MAO-A and MAO-B and the relative effectiveness of the R and S epimers were low. Therefore, contrary to expectations in the field,
-MPAI is useless as a drug.

The data presented below demonstrate that the high selectivity for one enantiomer over the other MAO is unpredictable. The structure of the MAO active site is not well understood to allow prediction of relative potency or selectivity of any given compound or pair of these enantiomers.

[0015]

[Table 1] One of the selective MAO-B inhibitors, (−)-deprenyl, has been extensively studied and is used as a MAO-B inhibitor to increase L-dopa therapy.
This treatment with (−)-deprenyl is generally effective, but does not cause a “cheese effect” at doses that result in near complete inhibition of MAO-B (Elsworth, et al., Psyc.
hopharmacology, 57, 33 (1978)). Furthermore, adding (-)-deprenyl to the combination of L-dopa and decarboxylase inhibitors administered to patients with Parkinson's disease improves akinesia and overall function, as well as on-off type variability. Associated with the removal of (review, Birk-m
ayer & Riederer in "Perkinson's Disease," Springer
-Verlag, pp. 138-149 (1983)). Thus, (-)-deprenyl potentiates, prolongs the effects of (a) L-dopa and does not increase the deleterious effects of L-dopa treatment.

However, (-)-deprenyl is itself
It has adverse side effects, including activation of preexisting gastric ulcers and occasional episodes of hypertension. Furthermore, (−)-deprenyl is an amphetamine derivative, and amphetamine and methamphetamine (these substances are metabolized to cause unwanted side effects such as increasing heart rate (Simpson, Biochemical Pharmacology, 27 , 1951 (197
8); Finberg, et al., In "Monoamine Oxidase Inhibit
ors-The State of the Art, "Youdim and Paykel, ed
S., Wiley, pp.31-43 (1981)).

There is a desire for other compounds which are selective irreversible inhibitors of MAO-B but which do not have the unwanted effects associated with (-)-deprenyl. As one of such compounds, N-propargyl-1-aminoindan.H
Cl (racemic PAI.HCl) is GB1,003
686 and GB 1,037,014 and 1970, 5
It is disclosed in U.S. Pat. No. 3,513,244 issued on May 19. Racemic PAI.HCl is a potent, selective irreversible inhibitor of MAO-B, is not metabolized to amphetamine and does not cause unwanted sympathomimetic effects.

Comparative animal studies have shown that racemic PAI has considerable benefit over (-)-deprenyl. For example, racemic PAI does not cause significant tachycardia, does not increase blood pressure (effect caused by a dose of (−)-deprenyl of 5 mg / kg), and doses of up to 5 mg / kg of nictitating membrane. It does not cause contraction or increase in heart rate (effect caused by (-)-deprenyl at a dose of 0.5 mg / kg or more). Furthermore, racemic PAI.HCl is
Does not enhance the cardiovascular effects of tyramine (Finberg, et a
l., in "Enzymes and Neurotransmitters in Mental Di
sease, "pp. 205-219 (1980), Usdin, et al., Eds., Wi
ley, New York; Finberg, et al. (1981), in "Monoami
ne Oxidase Inhibitors-The State of the Art, " ibi
d .; Finbergand Youdim, British Journal Pharmacol.,
85 , 451 (1985)).

One of the fundamental objects of the present invention is to separate racemic PAI compounds, and to enantiomers with MAO inhibitory activity, without any unwanted side effects associated with other enantiomers. It is to get what you don't.

Deprenyl has a structure similar to PAI, and the (-)-enantiomer of deprenyl, ie (-).
Since -deprenyl is known to be considerably more pharmaceutically active than the (+)-enantiomer, it would be expected that the (-)-enantiomer of PAI would be the more active MAO-B inhibitor. .

Contrary to this expectation, however, the (+)-PAI enantiomer is actually the active MAO-B inhibitor, but the (-)-enantiomer is very low.
It has been found to exhibit AO-B inhibitory activity. Furthermore,
The (+)-PAI enantiomer also has a surprisingly higher degree of selectivity for MAO-B inhibition than that of the corresponding racemic form, with less unwanted side effects than the racemic mixture in treating the indicated diseases. I won't. These findings are discussed in more detail below in vit
Based on both ro and in vivo experiments.

It has been subsequently shown that (+)-PAI has the absolute configuration of R. This finding is also surprising based on the predicted structural similarities of the (+)-PAI analogs with deprenyl and amphetamine.

As will be discussed below, [R] (+) PAI
Also of note is the high stereoselectivity of the pharmacological activity between the and S (-) enantiomers. Compound [R]
(+) PAI is almost four orders of magnitude more active than the S (-) enantiomer in inhibiting MAO-B. This ratio is significantly higher than that observed between the two deprenyl enantiomers (Knoll and Magya
r, Adv. Biochem. Psychopharmacol., 5 , 393 (1972);
Magyar, et al., Acta Physiol. Acad. Sci. Hung., 3
2 , 377 (1967)). Furthermore, in some physiological tests,
(+)-Deprenyl has been reported to have activity equal to or even higher than that of the (-) enantiomer (Tekes, et al., Pol. J. Pharmac.
ol. Pharm., 40,653 (1988)).

MPAI is a stronger inhibitor of MAO activity but has a lower selectivity for MAO-B than MAO-A (Tipton, et al., Biochem. Pharmacol., 31, 125).
0 (1982)). Surprisingly, the remarkable behavior of (R) (+) PAI is further clarified as only a small degree of difference in the relative activities of the two determined enantiomers was observed in MPAI (Table IB).

The object of the present invention is also to provide Parkinsonism, memory impairment, dementia, depression, hyperkinetic syndrome, emotional illness, neurodegenerative disease, neurotoxic injury, cerebral ischemia, traumatic injury to the head and spinal cord. A pharmaceutically active PA for the treatment of traumatic injury, schizophrenia, attention deficit, multiple sclerosis, or withdrawal symptoms
I-enantiomer alone (without L-dopa)
To provide a method to use (Youdim, et al.,
Review by, in Handbookof Experimental Pharmacolog
y, Trendelenberg and Wiener, eds., 90 / I, ch.3 (198
See 8)).

The present invention further provides a method of using the pharmaceutically active PAI-enantiomer alone for the treatment of Parkinson's disease. The present invention also provides [R] (+) P
There is provided a pharmaceutical composition containing a synergistic drug such as AI and levodopa. The use of such agents is effective when administered alone to patients with early stage Parkinson's disease and also has a synergistic effect on these patients when administered together with α-tocopherol, a vitamin E derivative. Has been studied for possible (−)-deprenyl (Th
e Perkinson's Study Group, New England J. Med., 32
1 (20), 1364-1371 (1989)).

In addition to its utility in treating Parkinson's disease, (-)-deprenyl also treats patients with Alzheimer-type dementia (DAT) (Ta).
riot, et al., Psycho-pharmacology, 91 , 489-495 (19
87)) and the treatment of depression (Mendelewicz and Youdim, Bri
t. J. Psychiat. 142 , 508-511 (1983)). The [R] (+) PAI compounds of the present invention and in particular their mesylate salts have been shown to restore memory. Thus, [R] (+) PAI has potential for treating memory disorders and dementia, especially the Alzheimer's version of children.

Finally, the present invention provides a very stable salt of [R] (+) PAI which has excellent pharmaceutical properties.
The mesylate salt was particularly stable, showing unexpectedly higher selectivities and significantly lower side effects than the corresponding racemic salt.

SUMMARY OF THE INVENTION The present invention provides R (+)-N-propargyl-1-aminoindan having the structure:

[0030]

[Chemical 1] The present invention further provides pharmaceutically acceptable salts of R (+)-N-propargyl-1-aminoindan.

The present invention further provides a therapeutically effective amount of R
Provided is a pharmaceutical composition containing (+)-N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The invention further provides a method of treating a patient afflicted with Parkinsonism, wherein an amount of R (+)-of the invention effective to treat Parkinsonism in the patient.
There is provided a method comprising administering to said patient N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof.

The present invention is a method of treating a patient suffering from memory impairment, wherein an amount of R (+)-N-propargyl-of the invention effective to treat the memory impairment of said patient. Further provided is a method comprising administering 1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method of treating a patient suffering from dementia, wherein an amount of R (+)-N-propargyl-1- of the present invention effective to treat the dementia of said patient. Further provided is a method comprising administering aminoindan or a pharmaceutically acceptable salt thereof to the patient. In one aspect, the dementia is of the Alzheimer type (DAT).

The present invention is a method for treating a patient suffering from depression, wherein an amount of R (+)-N-propargyl-of the present invention effective to treat the depression in the patient.
Further provided is a method comprising administering 1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient afflicted with hyperactivity syndrome, wherein an amount of R (+) of the present invention effective to treat hyperactivity syndrome in the patient. )
Further provided is a method comprising administering -N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient afflicted with an emotional illness, wherein an amount of R (+)-of the present invention effective to treat the emotional illness of said patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method of treating a patient suffering from a neurodegenerative disease, wherein an amount of the R (+)-N-of the present invention effective for treating the neurodegenerative disease of the patient. Further provided is a method comprising administering propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient afflicted with a neurotoxic injury, wherein an amount of R (+)-of the present invention effective to treat the neurotoxic injury in the patient. N
-Propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof is further provided to the patient.

The present invention is a method for treating a patient suffering from cerebral ischemia, wherein an amount of R (+)-of the present invention effective for treating cerebral ischemia in the patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient suffering from a traumatic head injury, wherein an effective amount of the R of the present invention for treating the traumatic head injury of said patient. (+)
Further provided is a method comprising administering -N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient suffering from a traumatic injury to the spinal cord, the method comprising: an effective amount of the present invention for treating a traumatic injury associated with the patient. R
Further provided is a method comprising administering to the patient (+)-N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof.

The present invention is a method for treating a patient suffering from schizophrenia, wherein an effective amount of R (+)-of the present invention for treating schizophrenia in said patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient suffering from attention deficit, wherein an amount of R (+) of the present invention effective to treat the attention deficit in said patient. ) -N
-Propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof is further provided to the patient.

The present invention is a method for treating a patient afflicted with multiple sclerosis, wherein an effective amount of the R (+)-of the present invention for treating multiple sclerosis in the patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method of preventing nerve damage in a patient, wherein an amount of R (+)-N-propargyl-1- of the present invention effective to prevent nerve damage in the patient. Further provided is a method comprising administering aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient suffering from withdrawal from an addictive substance, wherein an amount of R (+) of the present invention effective to treat the withdrawal of said patient.
Further provided is a method comprising administering -N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention provides R (+)-N-propargyl-
A method for producing 1-aminoindane, wherein R (-)-aminoindane is contacted with propargyl bromide or propargyl chloride or propargyl benzene sulfonate in the presence of an organic or inorganic base, whereby R ( Further provided is a method comprising forming a +)-N-propargyl-1-aminoindan and isolating the R (+)-N-propargyl-1-aminoindan thus formed.

The present invention relates to racemic N-propargyl-
A process for the preparation of 1-aminoindane, which comprises contacting racemic 1-aminoindane with propargyl bromide or propargyl chloride in the presence of an organic or inorganic base, whereby racemic N-propargyl-
Further provided is a method comprising forming 1-aminoindan and isolating the racemic N-propargyl-1-aminoindan thus formed.

Finally, the present invention is a process for the preparation of R (+)-N-propargyl-1-aminoindan salt, which comprises racemic N-propargyl-1-aminoindan as an optically active acid. To form two diastereomeric N-propargyl-1-aminoindan salts, and the diastereomeric N-propargyl-1-aminoindan salt thus formed is
Further provided is a method comprising isolating a (+)-N-propargyl-1-aminoindan salt.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides R (+)-N-propargyl-1 having the structure:
-Providing aminoindan.

[0052]

[Chemical 2] As shown in the following experimental example, [R] (+) PAI
Has an activity of [S] (−) P as an inhibitor of MAO-B.
Almost 7,000 times the AI. MAO-A and MAO
Considering MAO-B inhibitors known in the art, which have a low selectivity between -B and which do not show a predictable tendency for strength as a function of the R or S configuration, [R] (+ ) PA
The selectivity and strength of I is unexpected.

[R] (+) PAI can be obtained by optical resolution of a racemic mixture of the R- and S-enantiomers of PAI. Such a division is described in J. Jacques, A.
Collet and S. Wilen, "Enantiomers, Racemates and
It can be accomplished by any conventional resolution method known to those skilled in the art, such as that disclosed in Resolutions, "Wiely, New York (1981). For example, the resolution is performed by preparative chromatography on a chiral column. Other examples of suitable resolution methods include tartaric acid, malic acid, mandelic acid,
Or forming a diastereomeric salt with a chiral acid such as an N-acetyl derivative of an amino acid such as N-acetylleucine and then recrystallizing to isolate the diastereomeric salt of the desired R enantiomer. Is. PA
Racemic mixtures of the R and S enantiomers of I are, for example, GB 1,003,676 and GB 1,037,01
Prepared as disclosed in 4. Racemic mixtures of PAI can also be prepared by reacting 1-chloroindane with propargylamine. Alternatively, this racemic mixture reacts propargylamine with 1-indanone to form the corresponding imine and then reduces the carbon-nitrogen double bond of the imine with a suitable reagent such as sodium borohydride. Can be prepared by

According to the present invention, the R enantiomer of PAI reacts with propargyl bromide or salt face propargyl bromide or propargyl benzene sulfonate in the presence of an organic or inorganic base and optionally a suitable solvent. Can also be prepared directly from the optically active R-enantiomer of 1-aminoindane.

Suitable organic or inorganic bases for use in the above reaction include, for example, triethylamine, pyridine, alkali metal carbonates, and bicarbonates.
If the reaction is carried out in the presence of a solvent, the solvent may be selected, for example, from toluene, methylene chloride and acetonitrile. One method of making [R] (+) PAI is to react R-1-aminoindan with propargyl chloride using potassium bicarbonate as the base and acetonitrile as the solvent.

The reaction of 1-aminoindane described above generally results in a mixture of unreacted primary amines, the desired secondary and tertiary amines, N, N-bispropargylamino products. The desired secondary amine, N-propargyl-1-aminoindan, can be isolated from this mixture by conventional separation methods including, for example, chromatography, distillation and selective extraction.

The starting material R-1-aminoindan can be prepared, for example, from Lawson and Rao, Biochemisty, 19 , 2133 (1
980), the methods of the references contained therein, and the methods of EP 235,590, which are known in the art.

R-1-aminoindane is also for example:
Resolution of racemic mixtures of R and S enantiomers, including formation of diastereomers with chiral acids, or J. Jacqu
It can also be prepared by any other known method as reported by Es, et al., ibid . Alternatively, R-1-aminoindan is prepared by reacting 1-indanone with an optically active amine and then converting the resulting imine carbon-nitrogen double bond to palladium carbon, trivalent platinum or Raney nickel. It can be prepared by reduction by hydrogenation on any suitable catalyst. Suitable optically active amines include, for example, one of the antipodes of phenethylamine, or
Included are esters of amino acids such as valine or phenylalanine. The benzylic N—C bond can be subsequently cleaved by hydrogenation under mild conditions.

An additional method for preparing R-1-aminoindane is the indan-1-one oxime ester as described above, provided that the alkyl portion of the ester contains an optically pure chiral center. .) Hydrogenation.
Others include carbon-like imines or oximes
Non-chiral derivatives of indan-1-one containing a nitrogen double bond can be reduced with chiral reducing agents such as lithium aluminum hydride and ephedrine complexes.

The present invention further provides pharmaceutically acceptable salts of R (+)-N-propargyl-1-aminoindan.

In the practice of the present invention, pharmaceutically acceptable salts include mesylate, maleate, fumarate, tartrate, hydrochloride, bromate, esylate, p-toluenesulfonate, benzoate, Includes, but is not limited to, acetates, phosphates and sulfates.

In one embodiment, the salt is a mesylate salt of R (+)-N-propargyl-1-aminoindan, R
(+)-N-propargyl-1-aminoindan esylate salt, and R (+)-N-propargyl-1-aminoindan sulfate salt selected from the group consisting of night.

As shown in the following experimental examples, the mesylate salt was very stable to thermal decomposition and unexpectedly showed superior selectivity to MAO-B over the racemic salt.

To prepare pharmaceutically acceptable acid addition salts of the compounds of [R] (+) PAI, the free base is added to
It is reacted with the desired acid in the conventional manner in the presence of a suitable solvent. Similarly, acid addition salts can be converted to the free base form in a known manner.

A preferred method of preparing the mesylate salt of [R] (+) PAI is to add (a) propargyl benzenesulfonate (or tosylate or mesylate) to a 15% aqueous solution of sodium oxide. b) stirring for 5 hours; (c) adding additional toluene and water; (d) separating the organic layer, washing with 10% sodium hydroxide and then diluting with water;
(E) adjusting the pH of the mixture to 3.2 with 10% aqueous sulfuric acid; (f) separating the aqueous layer and adjusting the pH to 7.3 with 10% sodium hydroxide; (g) adjusting the pH. Extracting 3 times with toluene while maintaining; (h) combining the furniture layers and concentrating under reduced pressure to obtain a yellow oil; (i)
Dissolving the oil and L-tartaric acid in isopropanol; (j) heating under reflux for 1 hour; (k) cooling to room temperature and collecting the precipitate by filtration; (l) crude di-
Propargylaminoindan tartrate was recrystallized from methanol / isopropanol (1: 1) to give di (R (+)
-N-propargyl-1-aminoindan) tartrate; (m) dissolving the tartrate and methanesulfonic acid in isopropanol and heating under reflux for 30 minutes; and (n) cooling to room temperature and precipitating. Included is collecting R (+)-N-propargyl-1-aminoindan.

The present invention further provides a therapeutically effective amount of R.
There is provided a pharmaceutical composition comprising (+)-N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. A "therapeutically effective amount" of R (+)-N-propargyl-1-aminoindan or pharmaceutically acceptable salt thereof can be determined according to methods well known to those skilled in the art.

Possible salts for use in such compositions include the hydrochloride, phosphate, maleate, fumarate, tartrate, mesylate, esylate, and sulphate salts.

These compositions may be administered orally, parenterally, rectally,
Alternatively, it may be prepared as a drug that can be administered transdermally.

In one aspect, the pharmaceutically acceptable carrier is a solid and the pharmaceutical composition is a tablet. A therapeutically effective amount can be an amount of about 0.1 mg to about 100 mg.
A therapeutically effective amount can also be from about 1 mg to about 10 mg.

The forms described for oral administration include tablets, compressed or coated pills, dragees, sachets.
s), hard or soft gelatin capsules, sublingual tablets, syrups and suspensions.

In another embodiment, the pharmaceutically acceptable carrier is a liquid and the pharmaceutical composition is an injectable solution.
A therapeutically effective amount is from about 0.1 mg / ml to about 100 mg / ml
Can be an amount of A therapeutically effective amount is also about 1 mg / ml
To about 10 mg / ml. In one aspect, the dose administered is 0.1 ml to 1.0 ml.

In yet another embodiment, the carrier is a gel,
The pharmaceutical composition is a suppository.

For parenteral administration, the present invention provides ampoules or vials containing an aqueous or non-aqueous solution or emulsion. For rectal administration, suppositories with hydrophilic or hydrophobic vehicles are provided. For topical application, suitable delivery systems well known in the art such as ointments and those that can be delivered transdermally are provided.

In a preferred embodiment, the pharmaceutically acceptable salt is the mesylate salt.

These compositions may be used alone in the treatment of the above mentioned diseases or else as an adjuvant to conventional L-dopa treatment, as in Parkinson's disease.

The active ingredient in the above composition, namely [R]
The preferred dose of (+) PAI is within the following range.
For oral or suppository prescriptions, 0.
1 to 100 mg should be taken daily, preferably 1 to 10 mg per unit dose should be taken daily. For injectable formulations, 0.1-10 per unit dose
0 mg / ml can be taken daily, preferably 1-10 mg / ml per unit dose is taken daily.

In one embodiment, the pharmaceutical composition further comprises a therapeutically effective amount of levodopa. In another aspect, the pharmaceutical composition further comprises an effective amount of a decarboxylase inhibitor.

The amount of decarboxylase administered in combination with [R] (+) PAI or a pharmaceutically acceptable salt thereof is an amount effective to ensure uptake of L-dopa in the patient. Is.

The decarboxylase inhibitor may be L-carbidopa. In one aspect, a therapeutically effective amount of R
(+)-N-Propargyl-1-aminoindan is about 0.1 mg to about 100 mg, a therapeutically effective amount of levodopa is about 50 mg to about 250 mg, and an effective amount of L-carbidopa is It is from 10 mg to about 25 mg.

The decarboxylase inhibitor may also be ben-serazide. In one aspect, the therapeutically effective amount of R (+)-N-propargyl-1-aminoindan is from about 0.1 mg to about 100 mg, and the therapeutically effective amount of levodopa is from about 50 mg to about. 250 mg, an effective amount of benzerazide is about 12.5 mg to about 50 mg.

The invention further provides a method of treating a patient afflicted with Parkinson's disease, wherein the amount of the R (+)-N-propargyl-1-aminoindan of the invention is effective for Parkinson's disease in the patient. Alternatively, there is provided a method comprising administering a pharmaceutically acceptable salt thereof.

The use of [R] (+) PAI was tested with dopamine agonists, bromocriptine, pergolid.
Methods of treating Parkinson's disease in combination with e), lisuride, as well as other medications such as catecholamine oxidase methyltransferase inhibitors are within the scope of the invention.

In a preferred embodiment, the pharmaceutically acceptable salt is the mesylate salt.

The administration includes oral administration, rectal administration, transdermal administration,
Alternatively, it includes parenteral administration.

In one aspect, the invention further comprises administering to the patient a therapeutically effective amount of levodopa. In another aspect, the method of the present invention further comprises administering to the patient an effective amount of a decarboxylase inhibitor.

The decarboxylase inhibitor may be L-carbidopa. Another decarboxylase inhibitor is benzerazide.

The present invention is a method of treating a patient suffering from memory deficits, wherein an amount of R (+)-N-propargyl-of the present invention effective to treat the memory deficits in the patient. Further provided is a method comprising administering 1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method of treating a patient suffering from dementia, wherein an amount of R (+)-N-propargyl-1- of the present invention effective for treating dementia in the patient. Further provided is a method comprising administering aminoindan or a pharmaceutically acceptable salt thereof to the patient. In one aspect, the dementia is of the Alzheimer type (DAT).

The present invention is a method for treating a patient suffering from depression, wherein an amount of R (+)-N-propargyl-of the present invention effective to treat the depression in said patient.
Further provided is a method comprising administering 1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient afflicted with hyperactivity syndrome, wherein an amount of R (+) of the present invention effective to treat hyperactivity syndrome in the patient. )
Further provided is a method comprising administering -N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

Administration includes oral administration, rectal administration, or parenteral administration.

The present invention is a method for treating a patient afflicted with an emotional illness, wherein an amount of R (+)-of the present invention effective to treat the emotional illness of said patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method of treating a patient suffering from a neurodegenerative disease, wherein an amount of the R (+)-N-of the present invention effective for treating the neurodegenerative disease of the patient. Further provided is a method comprising administering propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient afflicted with a neurotoxic injury, wherein an amount of R (+)-of the present invention effective to treat the neurotoxic injury in the patient. N
-Propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof is further provided to the patient.

The present invention is a method for treating a patient suffering from cerebral ischemia, wherein an amount of R (+)-of the present invention effective for treating cerebral ischemia in the patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient afflicted with a traumatic head injury, wherein an amount of R of the present invention effective to treat the traumatic head injury of said patient. (+)
Further provided is a method comprising administering -N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient suffering from a traumatic injury to the spinal cord, wherein the amount of the invention is effective to treat the traumatic injury associated with the patient. R
Further provided is a method comprising administering to the patient (+)-N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof.

The present invention is a method for treating a patient suffering from schizophrenia, wherein an effective amount of R (+)-of the present invention for treating schizophrenia in said patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method for treating a patient suffering from attention deficit, wherein an amount of R (+) of the present invention effective to treat the attention deficit in said patient. ) -N
-Propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof is further provided to the patient.

The present invention is a method for treating a patient afflicted with multiple sclerosis, wherein an effective amount of R (+)-of the present invention for treating multiple sclerosis in said patient. Further provided is a method comprising administering N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The present invention is a method of preventing nerve damage in a patient, wherein the amount of R (+)-N-propargyl-1- of the present invention effective to prevent nerve damage in the patient. Further provided is a method comprising administering aminoindan or a pharmaceutically acceptable salt thereof to the patient.

In one aspect, the nerve injury is a structural nerve injury. In another aspect, the structural nerve damage is optic nerve damage.

The present invention is a method for treating a patient suffering from withdrawal from an addictive substance, wherein an amount of R (+) of the present invention effective to treat the withdrawal of said patient.
Further provided is a method comprising administering -N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof to the patient.

The term "withdrawal symptoms" as used herein refers to drug appeal, depression, hypersensitivity, anergy, amotivation (amotiv).
ation), changes in appetite, nausea, tremors and irregularities in sleep and physical and / or psycho-psychological symptoms.

As used herein, the term "addictive substance" refers to
For example, (a) addictive opiates such as opium, heroin and morphine, (b) psychostimulants such as cocaine, amphetamine and methamphetamine, (c) alcohol, (d) nicotine, (e) barbiturates, And (f) sedatives such as fentanyl, codeine, diphenoxylate and thebaine.

In one aspect, the addictive substance is cocaine.
In another aspect, the addictive substance is alcohol.

The present invention provides R (+)-N-propargyl-
A method for producing 1-aminoindan, comprising contacting R (-)-aminoindan with propargyl bromide or propargyl chloride or propargyl benzenesulfonate in the presence of an organic or inorganic base, whereby R (-) Further provided is a method comprising forming a +)-N-propargyl-1-aminoindan and isolating the R (+)-N-propargyl-1-aminoindan thus formed.

The present invention provides racemic N-propargyl-
A process for the preparation of 1-aminoindane, which comprises contacting racemic 1-aminoindane with propargyl bromide or propargyl chloride in the presence of an organic or inorganic base, whereby racemic N-propargyl-
Further provided is a method comprising forming 1-aminoindan and isolating the racemic N-propargyl-1-aminoindan thus formed.

Finally, the present invention is a method for preparing an R (+)-N-propargyl-1-aminoindan salt, which comprises adding racemic N-propargyl-1-aminoindane to an optically active acid. To form two diastereomeric N-propargyl-1-aminoindan salts, and the diastereomeric N-propargyl-1-aminoindan salt thus formed is
Further provided is a method comprising isolating a (+)-N-propargyl-1-aminoindan salt.

In one aspect, the isolation is by fractional crystallization
It includes isolating.

The following experimental details are provided to aid the understanding of the present invention and are not intended to limit the invention in any way as set forth in the claims that follow this specification, and Should not be understood as.

Experimental Details Example 1 Racemic N-Propargyl-1-aminoindane Hydrochloride
1-aminoindan and 10 of the racemate of a salt 10.0 g.
4 g of potassium carbonate was added to 75 ml of acetonitrile. The resulting suspension was heated to 60 ° C. and 4.5 g propargyl chloride was added dropwise.

The mixture was stirred at 60 ° C. for 16 hours, after which time most volatiles were removed by vacuum distillation. The residue was partitioned between 10% aqueous sodium hydroxide solution and methylene chloride.

The organic layer was dried and the solvent was removed by distillation. The residue was subjected to flash chromatography on silica gel eluting with 40% ethyl acetate / 60% hexane. Fractions containing the title compound as the free base were combined and the eluent replaced with ether. The ether solution is treated with HCl gas and the precipitate formed is isolated by suction filtration and recrystallized from isopropanol.
3 g of the title compound was obtained. m. p. = 182-4 [deg.] C. Chromatographic and spectral data are reported in U.S. Pat. No. 3,513,24 issued Mar. 19, 1970.
4, and in agreement with the reference sample were as follows.

NMR δ (CDCl ₃ ): 2.45 (2H, m), 2.60
(1H, t), 2.90 (1H, m), 3.45 (1H, m), 3.70 (2H,
d), 4.95 (1H, t), 7.5 (4H, m) ppm.

Example 2 S-(-)-N-propargyl-1-aminoindan salt
The Jo Compounds in the form of a salt free base, preparative HPL in Chiracel eluting with 10% isopropanol / 90% hexane OJ (cellulose tris [p- methylbenzoate])
On the C column, the racemic mixture of free base of Example 1 was resolved and isolated by collecting the first eluting major peak. The resulting oil was converted to the title compound by treating a 10% solution of the oil in diethyl ether with HCl gas and the resulting precipitate was collected by suction filtration. [Α] _D −29.2 ° (1%, ethanol), m.p.
p. = 182-184C. Other chromatographic and spectral properties were consistent with the hydrochloride salt of Example 1.

Example 3 R (+)-N-propargyl-1-aminoindan hydrochloride
Salt The title compound was prepared as in Example 2 above except collecting the second eluting peak from the preparative HPLC. [Α] _D
+ 29.1 ° (0.8%, ethanol), m.p. p. = 1
79-181 ° C. Other chromatographic and spectral properties were consistent with the hydrochloride salt of Example 1.

Example 4 R (+)-N-propargyl-1-aminoindan hydrochloride
Salt 12.4 g R (-)-1-aminoindan and 12.
9 g potassium carbonate was added to 95 ml acetonitrile. The resulting suspension was heated to 60 ° C. and 5.6 g of propargyl chloride was added dropwise. The mixture was stirred at 60 ° C. for 16 hours, after which time most volatiles were removed by vacuum distillation. The residue was partitioned between 10% aqueous sodium hydroxide solution and methylene chloride.

The organic layer was dried and the solvent was removed under reduced pressure. The residue was subjected to flash chromatography on silica gel eluting with 40% ethyl acetate / 60% hexane. Fractions containing the title compound as the free base were combined and the solvent replaced with ether. The ether solution was treated with HCl gas and the precipitate formed was isolated by suction filtration and recrystallized from isopropanol to give 6.8 g.
The title compound was obtained. m. p. = 183-185 ° C.
[Α] _D + 30.90 ° (2%, ethanol). The spectral characteristics were in agreement with those reported for the compound of Example 1.

Example 5 S (-)-N-propargyl-1-aminoindan hydrochloride
Salt Prepared by the method of Example 4 except using S (+)-1-aminoindan as the starting material. The product is [α]
_D -30.3 ° (2%, ethanol), m.p. p. = 18
3-5 ° C. The spectral characteristics were in agreement with those reported for the compound of Example 1.

Example 6A Di (R (+)-N-propargyl-1-aminoinda
L-tartrate salt To 48 ml of boiling methanol solution of tartaric acid (4.4 g) was added a solution of R (+)-N-propargyl-1-aminoindan free base (5.0 g) in methanol (48 ml). It was The solution was heated to reflux and 284 ml of t-butyl methyl ether was added over 20. The mixture was heated for a further 30 minutes, cooled and the resulting precipitate isolated by suction filtration to give 6.7g of the title compound. m. p. =
175-177 ° C .; [α] _D (1.5, H ₂ O) = + 3
4.3 °.

Elemental analysis: Calculated value for C ₂₈ H ₃₂ O ₆ N ₂ : C, 68.26, H, 6.56, N, 5.69. Found value: C, 68.76;
. H, 6.57; N, 5.61 Example 6B R (+) - N- propargyl-1-aminoindan main
To a solution of silate a) propargyl benzenesulfonate (78.4 g) and racemic aminoindane (63.2 g) in toluene (240 mL), a 15% aqueous sodium hydroxide solution (108 mL) was added dropwise at 20 ° C. After stirring for 5 hours, additional toluene (80 mL) and water (200 mL) were added with stirring. The organic layer was separated and washed with 10% sodium hydroxide then water. The pH of this mixture was adjusted to 3.2 by adding 10% aqueous sulfuric acid solution. Separate the aqueous layer, adjust the pH to 7.3 with 10% sodium hydroxide, p
Extracted three times with toluene, keeping H constant. The organic layers were combined and concentrated under reduced pressure to give 40.7 g of a yellow oil.

B) The above-mentioned crude racemic propargylaminoindan and L-tartaric acid (10 g) were dissolved in isopropanol (1 L), and the mixture was heated under reflux for 1 hr. The reaction was then cooled to room temperature with stirring and the precipitate was collected by filtration. The crude di-propargyl aminoindan tartrate salt was recrystallized from 1 L of 1: 1 methanol / isopropanol to give di (R (+)-N-propargyl-1-aminoindan) -L-tartrate salt. Physical and spectral data were consistent with that of the compound of Example 6A.

C) Di (R (+)-N-propargyl-1
-Aminoindan) -L-tartrate (15 g) and methanesulfonic acid (6 g) in isopropanol (150 mL)
The solution was heated to reflux for 30 minutes. The reaction was cooled to room temperature and the resulting precipitate was isolated by suction filtration and the title compound (11.
1 g) was obtained. This is the m. p. = 157 ° C. and [α]
_{With D} = 22 °.

Example 7 R (+)-N-propargyl-1-aminoindane hydrochloride
The form of the free base obtained in salt Example 4 R (+) - N- propargyl-1-aminoindan (1.2 g), potassium carbonate (0.97 g) and methyl iodide (1 g) in acetone 15mL In addition, the resulting suspension was heated to reflux under a nitrogen atmosphere for 8 hours. After this time, volatiles were removed under reduced pressure and the residue was partitioned between 10% sodium hydroxide (30 ml) and methylene chloride (30 ml). The organic layer was dried and the solvent removed under reduced pressure. The residue was subjected to flash chromatography on silica gel eluting with 40% ethyl acetate / 60% hexane. Fractions containing the title compound as the free base were combined and the solvent replaced with ether. The ether solution was treated with HCl gas. The volatiles were removed under reduced pressure and the residue was recrystallized from isopropanol to give 400 mg of the title compound as a white crystalline solid. m. p. = 134 to 136 ° C, [α] _D +31.4
0 (ethanol). NMR δ (CDCl ₃ ): 2.55 (2H, m);
2.7 (1H, br.s); 2.8 (3H, s); 3.0 (1H, m); 3.4 (1H,
m); 3.9 (2H, br.s); 5.05 (1H, m); 7.7 (4H, m) pp
m. Example 8 S (-)-N-methyl-N-propargyl-1-amino
Indane Hydrochloride The title compound was prepared in the same manner as in Example 7 above, except that S (−)-N-propargyl-1-aminoindan (free base) obtained in Example 5 was used as the starting material. The physical and spectral properties of the title compound are [α] D-34.9 °.
All were identical to that of Example 7 except (ethanol).

Example 9 Tablet composition N-propargyl-1 (R) -aminoindan hydrochloride 7.81 mg * Pregelatinized starch NF 47.0 mg Lactose NF hydrated 66.0 mg Microcrystalline cellulose NF 20.0 mg Sodium starch glycolate NF 2.99 mg Talc USP 1.5 mg Magnesium stearate NF 0.7 mg * Equivalent to 5.0 mg of N-propargyl aminoindan base.

Example 10 Tablet composition N-propargyl-1 (R) -aminoindan hydrochloride 1.56 mg * lactose hydrous 50.0 mg pregelatinized starch 36.0 mg microcrystalline cellulose 14.0 mg sodium starch glycolate 2.14 mg talc USP 1.0 mg Magnesium stearate NF 0.5 mg * Equivalent to 1.0 mg of N-propargyl aminoindan base.

Example 11 Capsule composition N-propargyl-1 (R) -aminoindan hydrochloride 5.0 mg Pregelatinized starch 10.0 mg Starch 44.0 mg Microcrystalline cellulose 25.0 mg Ethylcellulose 1.0 mg Talc 1.5 mg Required for granulation Pure water was added.

Example 12 Injectable composition N-propargyl-1 (R) -aminoindan hydrochloride 5.0 mg dextrose anhydrous 44.0 mg HCl, pH up to 5 1 ml was added with the necessary pure water.

Example 13 Injectable composition N-propargyl-1 (R) -aminoindan hydrochloride 1.0 mg sodium chloride 8.9 mg HCl, pH 5 was added to 1 ml with the necessary pure water.

Example 14 Injectable Composition N-Propargyl-1 (R) -aminoindan hydrochloride 2.0 mg Sodium chloride 8.9 mg HCl, pH 5 was added to 1 ml with the necessary pure water.

Example 15 Syrup composition N-propargyl-1 (R) -aminoindan hydrochloride 5.0 mg sucrose 2250.0 mg saccharin sodium 5.0 mg methylparaben 6.0 mg propylparaben 1.0 mg fragrance 20.0 mg glycerin USP 500 mg alcohol 95% USP 200 mg 5.0 ml pure water example 16 sublingual N- propargyl -1 (R) required - aminoindan hydrochloride 2.5mg microcrystalline cellulose 20.0mg lactose hydrous 5.0mg pregelatinized starch 3.0mg povidone 0.3mg colorant qs perfume qs Sweetener qs Talc 0.3 mg Excipient and active ingredient were mixed and granulated with povidone in ethanol. After drying and weighing, it was mixed with talc and compressed.

Example 17 PAI Sublingual Tablet N-Propargyl-1 (R) -aminoindan hydrochloride 5.0 mg Microcrystalline cellulose 15.0 mg Pregelatinized starch 12.0 mg Ethylcellulose 0.3 mg Talc 0.3 mg Necessary for granulation Pure water was added.

Example 18 Tablet composition N-propargyl-1 (R) -aminoindan hydrochloride 5.0 mg levodopa 100.0 mg carbidopa 25.0 mg pregelatinized starch 24.0 mg starch 40.0 mg microcrystalline cellulose 49.5 mg Col. D & C Yellow No. 10 0.5 mg Col. D & C Yellow No. 6 0.02 mg The alcohol USP required for granulation was added.

Example 19 Tablet Composition N-Propargyl-1 (R) -aminoindan mesylate 7.81 mg * Pregelatinized starch NF 47.0 mg Lactose NF water-containing 66.0 mg Microcrystalline cellulose NF 20.0 mg Sodium starch glycolate NF 2.99 mg Talc USP 1.5 mg Sodium stearate NF 0.7 mg * Equivalent to 5.0 mg of N-propargyl aminoindan base.

Example 20 Tablet composition N-propargyl-1 (R) -aminoindan mesylate 1.56 mg * lactose hydrous 50.0 mg pregelatinized starch 36.0 mg microcrystalline cellulose 14.0 mg sodium starch glycolate 2.14 mg talc USP 1.0 mg Sodium stearate NF 0.5 mg * Equivalent to 1.0 mg of N-propargyl aminoindan base.

Example 21 Capsule composition N-Propargyl-1 (R) -aminoindan mesylate 5.0 mg Pregelatinized starch 10.0 mg Starch 44.0 mg Microcrystalline cellulose 25.0 mg Ethylcellulose 1.0 mg Talc 1.5 mg Required for granulation Pure water was added.

The following examples and the accompanying tables and figures relate to the biological tests carried out according to the invention.

Example 22 In Vitro Inhibition of MAO Activity Experimental Protocol The MAO enzyme source is rat brain 0.3 M sucrose homogenate. This was centrifuged at 600 g for 15 minutes. Add the supernatant to 0.
Appropriately diluted in 05M phosphate buffer, preincubated with a dilution of a series of compounds ([R] (+) PAI, [S] (−) PAI and racemic PAI) for 20 minutes at 37 ° C. Next, a substrate labeled with ¹⁴ C (2-phenylethylamine (hereinafter referred to as PEA); 5-hydroxytryptamine, hereinafter referred to as 5-HT)).
And incubation for another 20 minutes (PEA),
Or it lasted for 30-45 minutes (5-HT). The concentrations of substrate used are 50 uM (PEA) and 1 mM (5-HT). In the case of PEA, the concentration of enzyme is chosen such that no more than 10% of the substrate is metabolized in the reaction pathway.
The reaction was then stopped by the addition of tranylcypromine (to a final concentration of 1 mM) and the incubation was filtered over a short column of Amberlite CG-50 buffered at pH 6.3. The column was washed with 1.5 ml of water, the eluate was stored and the content of radioactivity was determined by liquid scintillation spectroscopy. Since the amine substrate is all retained on the column, the radioactivity of the eluent indicates the neutral and acidic metabolites formed as a result of MAO activity. The activity of MAO in the samples was expressed as a percentage of the activity of the control in the absence of inhibitor after subtracting the appropriate blank values. PE as a substrate
The activity determined using A is designated MAO-B,
The activity determined using HT is called MAO-A.

Results The inhibitory activity of [R] (+) PAI, [S] (−) PAI and racemic PAI were tested separately in vitro and the results of a typical experiment are shown in FIGS. It was All experiments are
Repeated 3 times. The concentration of inhibitor resulting in 50% inhibition of substrate metabolism (IC-50) was calculated from the inhibition curves and is reported in Table 1.
Shown in B. From this data, the following can be seen.

(A) [R] (+) PAI is twice as active as the racemate for inhibition of MAO-B.

(B) [R] (+) PAI is 29-fold more active at inhibiting MAO-B than MAO-A.

(C) [S] (-) PAI is MAO-B
Only 1 / of the activity of [R] (+) PAI on the inhibition of
6,800, indicating low or no selectivity between MAO-B and MAO-A.

[0144]

[Table 1A] The results of a similar experiment using R (+) and S (-) MPAI (N-methyl-N-propargyl-1-aminoindan) are shown in Table 1B. The enantiomers of each MPAI are MAO-A and MAO-B rather than [R] (+) PAI.
The selectivity was low in the inhibition of. Furthermore, [R] (+)
MPAI [S] (−) MP for MAO-B inhibition
It was 5 times more active than AI. [R] (+) PAI
In contrast, [R] (+) PAI was approximately 7000 times more active than [S] (−) PAI in this assay.

[0145]

[Table 1B] Some experiments were also performed on human cerebral cortex tissue obtained 6 hours post-mortem and treated as described above. The results of such an experiment are shown in FIG. However, [R] (+) PAI, [S]
(−) PAI and racemic PAI are as defined herein.

Example 23 In Vivo Inhibition of MAO Activity: Short-Term Treatment Experimental Protocol Rats (from male Sprague-Dawley) weighing 250 ± 20 g were injected intraperitoneally (ip) or orally (oral gavage).
(Po) was treated with one of the enantiomers of PAI or the racemate and decapitated after 1 and 2 hours, respectively. Groups of 3 rats were used for each dose level of inhibitor and MAO activity in the brain and liver was determined using the general technique shown above. The amount of protein within each incubation was determined using the Folin-Lowry method and enzyme activity was calculated as nmol of metabolized substrate per time of incubation for each mg of protein. MAO in tissues obtained from animals treated with inhibitors
Activity was expressed as a percentage of enzyme activity in a group of control animals treated with vehicle (water for oral administration, 0.9% saline for ip injection) and killed as described above. expressed.

Results No significant change in behavior was produced at any dose level used in the inhibitor drug. The results are shown in FIGS. 4 to 11. i. p. After administration, [R]
(+) PAI is MAO-B in the brain at a dose of 0.5 mg / kg
Inhibition of 90% of The same dose is only 20% MA
It only resulted in the inhibition of OA activity. With oral administration, the same dose of [R] (+) PAI is equivalent to MAO-B.
Which resulted in 80% inhibition of MAO-A but no detectable inhibition of MAO-A. Substantially similar results are seen for inhibition of hepatic MAO as well as for brain MAO. The dose resulting in 50% inhibition of MAO-A and MAO-B (IC-50) was calculated from the inhibition curves and is reported in Table 2.
It was shown to. These data are (a) [R] (+) PA
The MAO inhibitory activity of I is maintained in vivo in rats; (b) [R] (+) in contrast to MAO-A
Selectivity for inhibition of MAO-B by PAI in vivo
maintained at o ; the very strong activity of the (+)-enantiomer in contrast to the (c) (-)-enantiomer
maintained in vivo; (d) the compound is effectively absorbed after oral administration; and (e) the compound effectively crosses the blood-brain barrier and effectively the brain MA.
It is shown to inhibit O. The fact that [R] (+) PAI is about 2-fold more active than the racemate for inhibiting MAO-B indicates that [S] (−) PAI is much less active for inhibiting MAO-B. Is a reflection of that.

[0148]

[Table 2] Example 24 Inhibition of MAO activity in vivo: Long-term treatment experimental protocol Rats (as specified in Example 23, 4 at each dose level) were orally dosed once daily for 21 days. Three dose levels (0.05, 0.1 and 0.5 mg / kg) were treated with [R] (+) PAI or racemic mixture and decapitated 2 hours after the last dose. MAO type A and B activity was determined in brain and liver as described in Example 23.

Results A daily dose of 0.1 mg / kg of compound [R] (+) PAI provided a good degree of selective inhibition.
This was 80% or more inhibition of brain MAO-B and 20% or less inhibition of brain MAO-A. At higher doses of 0.5 mg / kg per day, M
AO-A was still less than 50% inhibition (Figures 12 and 13). Liver MAO showed a similar degree of selective inhibition (Figures 14 and 15). The compound [R] (+) PAI was more potent than the racemic mixture due to a factor of approximately twice more. In case of brain MAO, [R] (+) PAI is MA
It had a better degree of selectivity for the inhibition of OB than the racemic mixture.

Example 25 Irreversible Experimental Protocol for MAO Inhibition A single dose of compound [R] (+) PAI (1 mg / kg) was administered i.p. p. Administered to a group of 4 rats by injection,
The animals were killed after 2, 6, 18, 24, 48 and 72 hours. MAO-B activity was determined in whole brain tissue as previously disclosed.

Results The results are shown in FIG. Maximum inhibition of MAO-B was reached 6 hours after injection. MAO activity only returned to 30% of control activity 72 hours after injection. This experiment demonstrates the irreversibility of inhibition of MAO by [R] (+) PAI.

Example 26 Intensity of the pressor effect of tyramine in conscious rats Experimental protocol Rats were anesthetized by intraperitoneal administration of a mixture of pentobarbital (30 mg / kg) and chloral hydrate (120 mg / kg). The left carotid artery and jugular vein were cannulated with a thin polythene tube (artery) or a thin silicone rubber tube (vein) connected to a polyethylene tube, the distal end of which was subcutaneously introduced into a fixed position behind the neck. The tube was viewed with heparinized saline and stoppered with a thin steel rod. Animals were treated with chloramphenicol by intramuscular injection and allowed to recover from surgery overnight. The next day, the rats were placed in a freely moving tall walled container. The intravenous catheter is connected to the pressure transducer via a 100 cm-long polyethylene tube with fine holes filled with physiological saline, and the intravenous catheter is connected to a 1 ml syringe via the same length tube. (The tube, including the syringe, contains an aqueous salt solution of tyramine hydrochloride (1 mg / ml)). After equilibration for 30 to 40 minutes, a tyramine injection (50 or 100 μg) was injected and the blood pressure response was recorded. At least a 15 minute interval was allowed between injections after blood pressure returned to control values. The pressure response of the control was confirmed, then 1 drug was injected intraperitoneally and the tyramine response was recorded over the next 4 hours. The area was evaluated according to the blood pressure response curve and the ratio of this area before treatment and after treatment to 1 to 3 hours after compound injection was calculated as
It was determined using the average of 3 to 4 values obtained during the control period.

Results The results are shown in Table 3. Compound [R] (+) PAI at a dose of 1 mg / kg, which causes complete inhibition of MAO-B in brain and liver and 40-50% inhibition of MAO-A in these tissues. Did not cause a sufficient increase in the pressor response of tyramine. A higher dose of [R] (+) PAI was 5 mg / kg (which was MAO-A in brain and peripheral).
Cause more extensive inhibition of. ), There was a significant increase in the pressor response of tyramine. This is, to a degree, similar to that provided by the same dose of deprenyl,
Lower than that induced by clorgyline (at doses that inhibit liver MAO-A activity by 85% or more).

[0154]

[Table 3] From this experiment, the compound [R] (+) PAI was converted to MAO-
It can be concluded that doses that effectively inhibit B do not cause an increase in the pressor effect of tyramine.

Example 27 MPAI-induced dopa with [R] (+) PAI
Suppression of minergic toxicity 1-Methyl-phenyl-1,2,3,6-tetrahydropyridine (MPTP) damages nigrostriatal dopaminergic neurons in several mammalian species including the mouse It is a neurotoxin that causes Parkinson's syndrome in humans and primates. An important early step in the mechanism of its neurotoxicity involves the conversion of MPTP to its toxic metabolite, 1-methyl-4-phenylpyridinium ion (MPP +). This reaction is based on the enzyme M
It is catalyzed by AO-B and probably occurs outside dopaminergic neurons, primarily in glia. MPTP
Is both a substrate for MAO-B and an irreversible inhibitor. Pretreatment of experimental animals with MAO-B inhibitors such as deprenyl or pargyline protects nigrostriatal neurons by blocking the oxidative conversion of MPTP to MPP +, thus reducing MPT.
P-induced damage to nigrostriatal neurons is prevented. Progressive nigrostriatal degeneration in Parkinson's disease results from exposure to environment-induced exogenous MPTP-like neurotoxins. In such cases, it is hoped that continued treatment with MAO-B inhibitors will mitigate the damaging effects of such yet putative MPTM toxicity, thereby slowing or slowing the progression of the disease. And from the very early stages of Parkinsonism
It is especially strongly pointed out that initiating continuous treatment with AO-B inhibitors. Effective MAO-B inhibitors are currently in
M of nigrostriatal dopaminergic neurons in vivo
It has been determined by the drug's ability to block PTP-induced damage. Therefore, the (-) and (+) enantiomers of PAI were tested for their ability to prevent or reduce MTPT-induced linear dopamine depletion in mice.

Experimental Protocol Male C57 black mice (20-25 g weight) were loaded with (a) M.
PTP-HCl (30 mg / kg dissolved in distilled water,
s. c. ) Or vehicle alone, or P
(-) Or (+) isomer of AI (2.5 mg / kg,
i. p. ) Or deprenyl (5 mg / kg, ip)
They were injected 1 hour after pretreatment with (b) and decapitated 5 days later. The brain was removed, the cadaver filaments were dissected on a glass plate cooled with ice and frozen on dry ice. An aliquot obtained by homogenizing linear tissue in 0.1 M perchloric acid and removing proteins containing dihydroxybenzylamine as an internal standard is dopamine and its main metabolites by electrochemical detection using HPLC. 3,4-dihydroxy-phenylacetic acid (DO
PAC).

Results Table 4 shows the results of this experiment. Treatment with MPTP resulted in a notable depletion of striatal dopamine (DA) and DOPAC. Treatment of PAI with the (−) and (+) enantiomers, or deprenyl did not affect the concentration of washed DA. Treatment of PAI with the (−) isomer gives MP
It did not affect TP-induced striatal DA and DOPAC levels. PA given before MPTP
The (+)-isomer of I completely abrogated the reduction of striatal DA and DOPAC levels caused by the toxin. At a dose of 2.5 mg / kg, the protective effect of (+) PAI was equivalent to that of (-) deprenyl (5 mg / kg).

[0158]

[Table 4] The above values for DA and DOPAC are mean ± SEM
And the number of rats in each group is n = 7-11.
Is.

These results show that [R] (+) PAI is
It is an excellent MAO-B inhibitor in vivo and has shown great potential for the treatment of Parkinson's disease.

The present invention is described in connection with, but not limited to, the above examples and the accompanying tables and figures.

Example 28 Amphetamine-induced Stem in Aged Rats
Effect of PAI Enantiomer on Rheotypic Behavior Amphetamine is known to induce stereotypic behavior by metabolism of endogenous dopamine (Sulser, F., and Sanders-Bush, E., Ann. Rev. Phar
macol., 11, 209-230 (1971)). Amphetamine is MA
Not metabolized by OB. Inhibition of MAO-B by administration of effective inhibitors and amphetamines resulted in inhibition of blocked MA
Causes the release of dopamine which is not degraded by OB. Therefore, the increase of synaptic dopamine leads to the increased efficacy of amphetamine and the effective MAO-, which leads to the increase of the stereotyped behavior under the influence of amphetamine.
Expected after administration of B inhibitor. A wide range of this behavior was evaluated by the number of lateral head movements per minute.

Experimental Protocol The test compounds were administered to drinking water at a dose of 0.5 mg / kg / day 24 hours before imposing hypoxia (92% nitrogen + 8% oxygen for 6 hours). After this, amphetamine 0.5 mg / kg
Sc at a dose of. After 45 minutes, lateral head movements were counted.

Results The results of these experiments are shown in Table 5.

[0164]

[Table 5] The results in Table 5 show that (+) PAI fully enhanced amphetamine-induced stereotypic behavior in both hypoxic-injured and control rats. (-)
PAI is totally inactive in this respect. these
The in vivo behavioral results reinforce previous biological findings that (+) PAI is an active inhibitor of MAO-B in the brain, but (-) PAI is inactive in this regard. Is.

Example 29 of [R] (+) PAI for memory improvement or recovery
Effects Newborn rat pups were subjected to short-term anoxia symptoms and then grown in the usual manner to develop long-lasting memory impairment (Speiser, et al., Behav. Br.
ain Res., 30, 89-94 (1988)). This memory impairment is manifested as poor performance in the passive avoidance test.

[R] for improvement or recovery of memory
The effects of (+) and [S] (-) PAI were tested in a passive avoidance test. If the drug is efficacious, it will prolong the latency of the response by going into the dark room or the room where the rat being tested has previously experienced an electric shock. The maximum response latency is 300 seconds.

Experimental Protocol Young rats were anesthetized postnatally as described in Example 27. [R] (+) PAI and [S] (−) P
AI was administered according to one of the following protocols.

Protocol A Mothers were given each isomer in drinking water at a dose of 1-1.5 mg / kg / day until weaning on day 21. After this, the weaned child was treated directly with the same dose for 20 days. Treatment 4
It was stopped on day 0 and the study was conducted on day 60, 20 days after the last dose of drug.

Protocol B dose was reduced to 0.5 mg / kg / day, administered to the foster mother on day 21 until weaning, and then directly to young rats until day 60 when the study was conducted.

Passive Avoidance Test The device consists of a bright room adjoining a dark room and a sliding door separating the two rooms. In training, the rat is placed in a bright room for 30 seconds and then the door is opened. The rat moves into the dark room during the incubation period and records this incubation period. When the rat enters the dark room, close the door and send a 0.3 mA foot shock for 3 seconds.

The test was repeated for recovery (memory) after 48 hours, and the incubation period of the step-through from the light to the dark was 30.
It was measured by recording against a temporary maximum of 0 seconds.

Results The results of these experiments are shown in Table 6.

[0173]

[Table 6] The experimental results show that (+) PAI, but not (−) PAI, is effective in improving memory in anoxic and control rats. The activity of the drug in this test is believed to be potentially effective in memory disorders, dementia, and senile dementia, especially of the Alzheimer type.

Example 30 Anoxia-induced hyperlocomotion in rat rats
Effects of [R] (+) PAI in the symptom group: Rats exposed to postnatal anoxia and then raised under normal conditions have increased exercise activity in the open place at 10-42 days of age. (Hertshkowitz, et
al., Dev. Brain Res., 7, 145-155 (1983)).

[R] (+) PAI and [S] (-) PA
The effect of I on such hyperkinetic syndrome was tested.

Experimental Protocol Rat pups were anoxic immediately after birth. Rat pups were placed in glass containers and exposed to 100% nitrogen for 25 minutes. The rat was resuscitated by gently applying an intermittent massage to the chest and then returned to its respective mother. Control rats were similarly treated with air instead of nitrogen.

[R] (+) PAI or [S] (-) PA
I (0.5 mg / kg / day) was administered to the foster mother in drinking water, thereby feeding the infant through breast milk. An electrical stimulus is transmitted across a 4 cm grid of infrared beams, which are sent to a measuring instrument. Exercise activity was recorded for 15 minutes at 15 and 20 days of age.

Results The experimental results are shown in Table 7.

[0179]

[Table 7] These results indicate that a high level of exercise was achieved by reaching the point where breast cancer was fed by long-term oral treatment with [R] (+) PAI at a dose of 0.5 mg / kg administered to the foster mother. Progressive syndrome is improved. Therefore, [R] (+) PAI
Is a potentially effective medicine for the treatment of hyperkinetic syndrome in children.

Example 31 Salt Stability Differences of Ten PAIs Stability is an important factor in selecting the optimal salt as a therapeutic agent. Various salts can change the physicochemical and biological properties of a drug, having a dramatic effect on its overall properties. (Berge. SM, et al., J. Pharm.
Sci. 66 , 1 (1977); Gould, PL, Int. J. Pharmaceu
tics, 33 , 201 (1986)).

Experimental Synthesis of PAI Salt A solution of the appropriate acid (1 mol equivalent) in 2-propanol was added to P
AI (1 mol equivalent) was added to a solution in 2-propanol with stirring (Ar, BHT). The salt formed is filtered,
It was washed with 2-propanol and ether and dried under low pressure. The yield was 70 to 90%. Except when producing PAI acetate, the use of ether as a solvent is included.

Analytical Method Lichrosphere 60 RP Select B 5m 125 × 4 mm (Merck) column, L-4200UV-Vi set at 210 nm.
HPLC equipped with s detector (Merck-Hitachi) (Jasco BI
P-1) and D-2500 chromatographic integrator (Merck-Hitachi) were used for chromatographic separation. The eluate and diluent were 80% distilled water / 20% acetonitrile (HPLC grade), and ammonia water
It consists of 0.07M perchloric acid adjusted to H2.5. The flow rate used was 1 ml / min, the concentration of the appropriate PAI salt solution was 250 μg / ml and 20 μl of the solution was injected into the chromatographic system.

The range of melting point is determined by an automatic device (Mettler FP 8
0), thermogravimetric analysis is applicable at 10 ° C.
Performed on a Mettler TA 3000 system at a rate of / min. Solubility was determined in appropriate dilutions of supernatants obtained from saturated aqueous solutions of PAI salts and measured on a UVIKON 941 (Kontron) UV-Vis spectrophotometer. The salt form (mono- or di-salt) is C,
Obtained by elemental analysis using standard equipment for determining H, N and S. The pH was measured with a 1% aqueous solution of PAI salt.

Results The properties of various salts are summarized in Table 8.

[0185]

[Table 8] Competitive stability tests were conducted under a series of several accelerated conditions. I) Heating at 80 ° C. for 72, 96 or 144 hours; II) Refluxing in isopropanol for 30 hours.
The generated decomposition product was measured by HPLC and confirmed by TLC. The results are shown in Table 9 as a percentage of the area over the total area of the integrated peak, together with the relative retention times (for PAI peak; RRT).

[0186]

[Table 9] The salt was subjected to visual inspection for color and shape. The findings are shown in Table 10.

[0187]

[Table 10] These tests show that sulphates, esylates and mesylates are sufficiently beneficial compared to other salts due to their good solubility and chemical stability. Of these three types of salts, mesylate is preferable because it has excellent stability even under destructive conditions.

Example 32 Haloperidol-induced catalepsy in mice
Over recovery male ICR mice were pretreated with either of the following drugs each 25-30 g: Saline, [R] (+) PAI mesylate, or racemic PAI mesylate. All drugs are
It was administered ip in a volume of 0.2 mL. Two hours later, haloperidol was injected sc in a volume of 0.1-0.2 mL at a dose of 6 mg / kg. Exercise coordination test 3 hours after administration of haloperidol, ie 5 minutes after administration of putative protective drug
Went in time.

The exercise coordination test and rigidity are three different factors: (a) a horizontal rod of a given length, the ability to walk a length of 80 cm; (b) a vertical rod with the face down, a length of 80 cm. Ability to crawl; (c) Quantification according to duration of rest in an unnaturally seated position in which the abdomen of the mouse is pressed against the "wall". Four points were given in each trial, or 12 points in all trials, for complete performance similar to mice not treated with haloperidol. Those with lower abilities gave 1 to 3 points. The key scores are shown in Table 9A. The effect of various agents on antagonizing haloperidol-induced catalepsy is shown in Table 11. 3 hours after administration of haloperidol, [R] (+) PAI mesylate was 5-15
7. Provides protection against haloperidol at mg / kg,
The peak of action is reached at 5 mg / kg (activity score = 94% of saline control). Racemic PAI mesylate conferred partial protection in the range of 7.5-15 mg / kg and was inactive at 5 mg / kg. From FIG. 17, [R]
The dose-effect profile of (+) PAI mesylate or racemic PAI may be such that an increased dose above 10 mg / kg is accompanied by a diminished effect, whereas the racemic mixture is generally less potent. Recognize. This means that the racemic PAI mesylate will always be less active than the (R) enantiomer at two doses of [R] (+) PAI mesylate.

Rats with α-MpT-induced hypolocomotion
, A recovery drug, α-MpT, is the formation of L-dopa from tyrosine
And as a result, it is postulated to inhibit the formation of dopamine itself. Lack of CNS dopamine manifests as hypoactivity. Six-month-old male Wistar rats (Harlan Orkack,
(Obtained from UK) were pretreated with saturated saline, [R] (+) PAI mesylate or RacPAI mesylate at the indicated doses. 2 hours later, the mouse was ip to α-Mp
T was administered at a dose of 100 mg / kg in 0.3-0.5 mL. After this, locomotor activity was recorded in an active cage with a computer for 10 hours. The results are shown in Table 1.
2 and FIG. [R] (+) PAI at 2 mg / kg
Mesylate is about 90% of rats treated with saturated saline
Restored the level of activity until. In each case, the dose-effect curve profile is bell-shaped, which is between 2 and
It is suggested that the effect decreases as the dose increases beyond the peak of 5 mg / kg. RacPAI at 5 mg / kg
Mesylate is unable to elicit a level of activity comparable to that of [R] (+) PAI mesylate at 2 mg / kg.

From these measurements, [R] (+) PAI mesylate and RacPAI mesylate do not show similar activity patterns in the recovery of normal motion sickness in mice treated with haloperidol and in rats treated with α-MpT. [R] (+) PA at all doses tested
I-mesylate is usually more potent than the corresponding dose of RacPAI mesylate. Also, at a given dose, RacPAI mesylate is always less effective than [R] (+) PAI mesylate at half the same dose. [R]
Doubling the dose of RacPAI mesylate over (+) PAI mesylate does not have the same effect as that of [R] (+) PAI mesylate.

Pharmacologically, consider that RacPAI mesylate consists of 50% of the active ingredient (which is [R] (+) PAI mesylate) and 50% of an inactive substance such as a diluent. I can't. The presence of [S] (-) PAI within the RacPAI mesylate indicates [R] (+) P
It has the opposite effect on the activity of AI, causing more than a 2-fold decrease in strength. This decrease may be due to the direct adverse effect of [S] (-) PAI on the behavioral parameters.

Table 11 Restoration of haloperidol-induced catalepsy in mice using [R] (+) PAI mesylate and racemic mesylate Mice were given each test drug at the dose indicated in ip. Two hours later they were given haloperidol as indicated in the text. The doses given are for the free base.

[0194]

[Table 11] Statistical advantage over haloperidol alone: Student's
* P ≦ 0.05; ** p ≦ 0.01 by “t” test;
*** p ≦ 0.001.

The score for [R] (+) PAI was significantly different from that for racemic PAI. This is 5
p ≦ 0.05 at mg / kg; p ≦ 0.01 at 10 mg / kg; and p ≦ 0.05 at 15 mg / kg.

[0196]

[Table 11A] Table 12 Restoration of locomotor activity in rats treated with α-methyl-p-tyrosine (α-MpT) at 100 mg / kg ip Rats were given the test drug at the dose indicated in ip.
Two hours later they were given haloperidol and quickly placed in the activity basket. Total locomotor activity was recorded automatically for 10 hours as indicated in the text.

[0197]

[Table 12] Student's “t” test statistical advantage: α-MpT
* P ≦ 0.01 vs. test drug alone + α-MpT;
*** p ≦ 0.001.

[R] (+) for racemic PAI
PAI scores were significantly different at 2 mg / kg.

Example 3 [R] (+) PAI in rats after covering the wound
Mesylate effect method 1. Induction of trauma Head trauma is represented by 1 to 2 in the left cerebral hemisphere, ie, central coronal plane
Male rats were induced under ether anesthesia by a weight drop device well calibrated to drop onto the exposed skull over the mm side to midline.

2. Assessment of Motor Function One hour after induction of trauma, the rats were tested by a series of criteria to assess these psychiatric outcomes (the criteria are Shohami, et al., J. Neurotrauma, 10 , 113 ( 199
3)). Psychiatric severity score (N
These criteria, called the SS; Neurological Severity Score, consist of a series of reflex and motor functions. Points are given below the lack of these criteria. Rats were reassessed at 24 hours.

3. Assessment of cerebral edema The brain was removed after the second assessment of motor function and a piece of tissue (-20 mg) was weighed to obtain wet weight (WW). 24
After drying in a desiccator oven at 95 ° C. for an hour, it was weighed again to obtain a dry weight (DW). The percentage of water in the tissue was calculated as (WW-DW) x 100 / WW.

4. Drug treatment [R] (+) PAI mesylate was dissolved in water and administered to rats at a dose of 0.1 mg / kg after induction of head trauma 0,4,
It was administered intraperitoneally at 8 and 12 hours. Control rats were treated with water for the same amount of time.

Results The NSS, which measures the "clinical status" of the rats, was almost identical in the treated and untreated groups 3 hours after head trauma, but was treated with [R] (+) PAI mesylate. In rats, there was a sufficient decrease at 24 hours (Table 13). These results indicate that PAI mesylate is effective in improving recovery of motor function after closing head injury in rats.

Twenty-four hours after trauma, major edema was found in the left hemisphere (78.5% water in intact brain tissue versus 85.4% water in control rat brain). PAI mesylate is
It is effective in reducing edema confirmed by its effect on water percentage.

As a result, the results reported here are
We show that [R] (+) PAI mesylates have neuroprotective properties in a model directed towards inducing trauma to the same human neurotrauma and the skull when closed.

[0206]

[Table 13] Example 34 To frost damage of NMDA-induced cell death of cerebellar cell culture
Effect of PAI Mesylate on Procedure: Mechanically Dissociated Neonatal Rat Cerebellar Cultures Aseptically dissociate cerebellum from 6 or 7 day old rat pups,
3 ml of enriched medium (high medium glucose concentration (1 g /
l) Dulbecco's containing 2 mM (v / v) L-glutamine, a cytostatic mixture of antibiotics and enriched with 15% (v / v) fetal calf serum inactivated. Modified Eagle medium (DMEM). ) Containing 15 ml
Placed in a sterile plastic conical tube. next,
The cerebellum was dissociated after 20 to 25 sterile 13 gauge 10 cm long stainless steel needles attached to a 5 ml syringe with a 45 μm pore size nylon sieve inserted. The dissociated cells were centrifuged at 200 g for 5 minutes, the supernatant discarded and the cells resuspended in enriched medium. The viability of the cells was determined by trypan blue exclusion test. The cells are then immersed in poly L-lysine coated surface (poly L-lysine coated glass coverslips in sterile distilled aqueous solution containing 15 μg / ml poly-L-lysine, Wash with sterile water immediately before use,
Prepared for at least 1 hour before plating by drying. ) At a density of 200 mm ² and covered with enriched medium, under an atmosphere of 5% CO _{2 in} air and 100%.
Incubated at 37 ° C in humidity. After 4 days of culture, the medium was replaced with medium containing the desired test compound. The experiment was done exactly the same and was repeated 2 or 3 times.
After determining the toxic dose-response of test compounds, the four groups were compared. (I) Control (enriched medium only), (II)
Test compound (1 subgroup for each concentration (two concentrations were tested)), (III) N-methyl-D-aspartate (NMDA, 1 mM for cytotoxic challenge)
Was exposed for 3 hours. ), (IV) Test compound and NM
DA (1 subgroup for each of the two concentrations of the test compound), (V) a control group for testing the effect of the solvent in which the test compound is dissolved, and (VI) Additional "positive control" groups of spermine (dissolved at 0.01 μM in culture medium) and NMDA. Neuronal viability was assessed after 24 hours by phase contrast microscopy and trypan blue staining.

Results Glutamate (Glu) has been shown to have several neuromedical disorders including epilepsy and seizures, and most suitably brain degeneration of the brain such as Parkinsonism, Alzheimer's disease and traumatic brain injury. It was well established to have neurotoxicity expressed in. The neurotoxic effect of Glu is the membrane-bound N-methyl-D-aspartate (NMDA).
It is mediated by Glu receptors such as receptors.

The results shown in Table 14 show that 10 μM [R] (+) PAI mesylate increases cerebellar cell viability by 27% after exposure to 1 μM NMDA. These in vitro results indicate the in vivo effect of [R] (+) PAI mesylate shown in Examples 33 and 35, in which the agent has neuroprotective properties against the neurotoxic concentration of NMDA. Indicates that it has.

[0209]

[Table 14] Values expressed as percentages of untreated controls represent the mean of two identical experiments for culture experiments and the mean ± SEM of 4 animals for collapse. The percent protection value is the effect of the test compound after subtracting the effect of the solvent.

Example 35 [R] (+) PAI Membrane Following Gradual Contusion of the Rat Optic Nerve
Effect of Sylates The neuroprotective effect of [R] (+) PAI mesylate was determined for rapid application after crush injury of the optic nerve of adult rats. Short-term effects were measured metabolically and long-term effects were measured electrophysiologically.

Method 1. Metabolism measurement a) General method is Yoles, et al., Investigative Ophthalmology
& Visual Science, 33, 3586-91 (1992). In the short term, metabolic measurements are monitored by the mitochondrial NADH / NAD ratio, which depends on the activity of the electron transport chain, which indicates the level of energy production. Changes in the nerve's ability to produce energy as a result of injury are determined by comparing the levels of NADH in response to artificial transient anoxic injury before and after injury.

B) Surface Fluorescence Assay-Refluorescence Assay The monitoring of the NADH redox status in mitochondria is based on the fact that, unlike the oxidized form NAD ⁺ , NADH fluoresces, at which time it emits light at 450 nm. . A flexible Y-shaped bundle of optical fibers (a light guide) was used to transmit light to and from the optic nerve. Light emitted from nerves has two wavelengths: 366 nm
(Reflected light) and 450 nm (fluorescence). Changes in reflected light were corrected by hemodynamic effects and changes in tissue absorption due to changes in the secondary optic nerve that alter arterial blood pressure and nerve volume. Fluorescence measurements have been found to be sufficiently accurate for measuring NADH redox state by subtracting the reflected light (366 nm) from the fluorescence (1: 1 ratio) and obtaining a corrected fluorescence signal. .

C) Animal Preparation Animal utilization refers to ARVO R for use of animals in research.
Followed esolution. Male Spra- weighing 300-400 g
gue-Dawley (SPD) rats were anesthetized with sodium pentabarbitone (50 mg / kg, ip). The animal's head was properly held by a head holder, lateral lateral canthal incision was made with binocular surgical microscopy, and the conjunctiva was incised lateral to the conjunctiva. After separating the posterior myoblasts, confirm the optic nerve, and roughly incise a length of 3 to 3.5 mm (bl
It was exposed near the eyeball. Care was taken to leave the dura intact and not damage the nerves. A special light guide holder was implanted around the optic nerve so that the light guide was located on the surface 1 mm distal to the optic nerve with respect to the injury site. While still anesthetized, animals were allowed to recover from surgery and then exposed to anoxic conditions. Anoxic status is 10
This was accomplished by breathing the rat for 2 minutes in an atmosphere of 0% nitrogen, then returning to air. To assess the metabolic activity of the optic nerve, relative changes in reflected light and fluorescence intensity in response to anoxia were measured before and after crushing.

D) Experimental protocol for contusion and metabolism measurements
Cole With the help of calibrated cross section forceps, a moderate bruise was applied to the optic nerve between the eye and the light guide holder for 30 seconds at a pressure corresponding to 120 g.
Immediately after injury, animals were injected ip with and without [R] (+) PAI mesylate (2 mg / kg). To assess the activity of the energy generating system, the NADH response to 2 minutes of anoxia,
All animals were measured before injury, 30 minutes after injury, and thereafter at 1 hour intervals for up to 4 hours (see Figure 19).

2. Electrophysiological measurements This method is based on Assia, et al., Brain Res., 476, 205-21.
2 (1989). The animal preparation and optic nerve damage are preferably similar to metabolic studies. Immediately after injury, animals received [R] (+) PAI mesylate (0.5
Water containing (mg / kg) and water without this were injected once. The optic nerve was excised 14 days after injury and treatment and measured electrophysiologically. Rats were deeply anesthetized with 70 mg / kg pentabarbitone prior to removal of the optic nerve for electrophysiological measurements. The skin was removed from the skull and the optic nerve was torn from the eye. A nearly total decapitation was performed and the skull was opened with rongeur. The cerebrum was replaced laterally to expose the intracranial portion of the optic nerve. The incision is at the level of the nerve, and the nerve is used as NaCl (126 mM), KCl (3 m
M), NaH ₂ PO ₄ (1.25 mM), NaHCO
₃ (26 mM), MgSO ₄ (2 mM), CaCl ₂ (2 m
M) and D-glucose (10 mM) in a fresh salt solution containing 95% O ₂ and 5% at room temperature.
Exposed to CO ₂ . The nerve was kept in this solution. Here, the electrical activity remains stable for at least 3-4 hours. 0.5 hours after collection at room temperature, electrophysiological recordings were obtained from the optic nerve distal to the contusion area. next,
Two aspirations of the nerve endings immersed in the immersion solution at 37 ° C
It was connected to a g-AgCl electrode. A stimulation pulse was applied from the electrode at the closest end and action potentials were recorded at the distal electrode. Gr
The ass SD9 stimulator was used for ultra-maximal electrical stimulation. The measured signal is transmitted to a Modelec PA36 preamplifier and then to an electromyography (Medelec MS7, AA7T amplifier). The maximum amplification of the action potentials (CAPs) of the eight averaged compounds was recorded and photographed with a Polaroid® camera.
CAP values were measured on the contralateral undamaged nerve, which served as a reference.

Results Results were applied rapidly after optic nerve injury [R].
(+) PAI mesylates have been shown to block injury-induced reduction in energy production. [R]
(+) PAI mesylate also has long-term effects as measured by electrophysiological monitors.

CAP (compound action potential, compound actio)
n potentials) directly correlated with the number of fibers processed in the tested part of the nerve.

[R] (+) PAI mesylate significantly alleviates injury-induced loss in the distal portion of the injured nerve. This means that [R] (+) PAI
Mesylate is a neuroprotective agent or at least degenerate
Has been shown to delay .

[0219]

[Table 15] Example 36 Antispasmodic salt of [R] (+) PAI and [S] (-) PAI
Comparison of seizure properties Both the [R] (+) PAI and [S] (−) PAI HCl salts have significant anticonvulsant activity. Mice (ip administration) in maximum electric shock test (MES test)
So, [S] (-) PAIHCl is [R] (+) PAIH
It had an anticonvulsant activity (ED ₅₀ = 57 mg / kg) greater than Cl (ED ₅₀ = 79 mg / kg). Similar results were observed in rats (po dose). 4 out of 4 rats
Animals were protected from seizures in the MES test when they were dosed with 50 mg / kg [S] (−) PAIHCl. On the other hand, after administration of the same amount of [R] (+) PAIHCl, 3 out of 4 mice were protected. With regard to its effect on Parkinson's disease, enhanced anticonvulsant activity is a deleterious side effect. A similar trend occurs with mesylate salts. [S]
(−) PAI mesylate was [R] (+) in MES test.
It has greater anticonvulsant activity than PAI mesylate. 1
At a dose of 00 mg / kg, [S] (-) PAI mesylate protected 3 out of 3 mice, while only 1 out of 3 mice [R] (+) PAI mesylate. Was defended by.

The MES test is the classic model for indicating efficacy against partial and systemic seizures in humans. The mechanism of action of drugs mediates their ability to prevent the spread of seizures. However, some drugs that prevent the spread of seizures have the side effect of lowering the seizure threshold.
Therefore, these drugs have both preconvulsant and anticonvulsant side effects.

The results here show that [S] (-) PAI mesylate has preconvulsant activity. 141 mg in the "Timed intravenous infusion study of metrazol"
[kg] / kg [S] (-) PAI mesylate reduces the time and therefore the amount of metrazole required to induce both the first focal seizure and the onset of clonus. Other agents classically used for partial and systemic seizures, such as phenytoin and carbamazepine, do not show this effect. (HJ Kupferberg, Epil
epsia, 30, s51-s56 (1989)). Similarly, [S] (-)
PAI mesylate exhibits significantly higher acute neurotoxicity than [R] (+) PAI mesylate. At 300 mg / kg,
[R] (+) PAI mesylate did not show any neurotoxicity in mice in the rotor rod ataxia test. [S] (-) PAI
With mesylates, 4 out of 4 mice were neurotoxic and convulsive.

Method TD50 (Central Toxic Dose) This test measures psychiatric deficits by the rotor rod ataxia test. Mice were placed on a knurled rod rotating at 6 rpm. Then, whether the mouse has the ability to maintain its equilibrium, and 1 in each of the 3 tests.
Decide if you can stay on the rod for a minute.

Time Intravenous Infusion Study of Metrazole This study measures the minimum seizure threshold in each animal. Metrazol was injected at 0.185 mg / ml into the tail vein of mice. The time was then recorded from the onset of infusion to the first twitch (first focal seizure) and clonus onset (clonic seizure). Preconvulsions require less metrazole to cause these signs and thus end in a shorter time.

Example 37 [R] (+) PAI and Contraction of Intestinal Smooth Muscle Preparation
Peripheral effect of [S] (−) PAI The peripheral effect of the hydrochloride of the enantiomer of PAI was determined in the isolated small intestine of rabbits or black-headed mice. These observations provide valuable information regarding their relative peripheral side effects in humans. The first point of contact between an orally administered drug and the patient is between the gastrointestinal tract, where the concentration of drug is higher than after absorption and distribution. PAI hydrochloride (M
In the case of W = 208), an oral dose of 10 mg contained in a liquid volume of about 100 ml equates to a concentration of about 0.5 mM. In contrast, therapeutic plasma concentrations of [R] (+) PAI hydrochloride are on the nanomolar order.

The effect of the enantiomers of PAI on the isolated rabbit jejunum and guinea pig ileum was determined and from this result [S] (−) PA together with [R] (+) PAI was determined.
It is to be seen whether uptake of I (as found in racemic PAI) results in side effects that are not present upon administration of pure [R] (+) PAI. [R] (+) PA
I is the preferred enantiomer for inhibition of MAO-B in the brain due to its potency and high selectivity for this form of the enzyme. [S] (−) PAI has a lower strength than [R] (+) PAI in this regard, and MA
It is also not selective for OB. Basically, [S]
Its presence in the racemic form of PAI is acceptable and overlooked, provided that (−) PAI is inactive at the recommended dose of [R] (+) PAI. The results shown in Tables 16 to 19 show that [S] (-) PAI is an inactive substance. In contrast, in the murine ileum, it is a stronger relaxant than [R] (+) PAI. Therefore, its erasing effect cannot be discounted and considered because it can be ignored. These data are pure [R]
The deleterious side effect of (+) PAI administration is [R] (+)
It is shown to be smaller than in the administration of racemic PAI containing a dose equivalent to PAI.

Table 16 P in jejunal preparations soaked in water
Enhancement of Tyramine by Each of the Two Enantiomers of AI Rabbit jejunum stretches were mounted in an organ bath and a periodic contraction observed by norepinephrine but not tyramine was observed. However, when the jejunum is pretreated with a monoamine oxidase inhibitor such as PAI, tyramine causes spontaneous contractile relaxation. The degree of relaxation can be correlated with the relative strength of the inhibitor.

[0227]

[Table 16] Results [S] (-) PAI is much less potent than [R] (+) PAI as an inhibitor of brain MAO-B. Therefore, [S] (−) PAI is not an effective drug for inhibiting the breakdown of brain dopamine, but it can enhance the tyramine-evoked norepinephrine release in the small intestine. Its activity in the small intestine is an unwanted side effect as it is expected to enhance the absorption and action of undegraded tyramine. Therefore, [S] (-) PAI is
When used with [R] (+) PAI, it is not an inactive material as found in racemic PAI.

Table 17 Antagonism of bethanechol-induced contraction of guinea pig ileal preparations in the presence of each of the two enantiomers of 400 μM PAIHCl, mounted in physiological solutions of organ baths. Ileal stretch contracts in a dose-dependent manner when treated with bethane-chol, an enzymatically stable analog of the natural gastrointestinal neurotransmitter acetylcholine. These contractions are weakened in the presence of PAI.
Data were expressed as gram-tension.

[0229]

[Table 17] The result [S] (-) PAI is MAO- with respect to [R] (+).
Almost inactive as a B inhibitor. Therefore, it is not effective in inhibiting the breakdown of brain dopamine.
However, it is more effective than [R] (+) PAI in inhibiting the small bowel contraction induced by bethanechol. Therefore, in [S] (-) PAI, this is [R]
Racemic PAI when used with (+) PAI
It is not an inactive substance as found in.

Table 18 Antagonism of histamine-induced contraction of the guinea pig ileal preparation by each of the two enantiomers of PAIHCl. A fixed dose of histamine (40 μM) was mounted in physiological solution of an organ bath. Causes a continuous contraction of the ileal stretched material of the swollen guinea pig. Increasing addition of each of the two enantiomers of PAIHCl causes a dose-dependent relaxation of muscle. Results were expressed as percent relaxation relative to baseline (this was taken as 100% relaxation) before addition of histamine.

[0231]

[Table 18] Results [S] (−) PAI is inactive with [R] (+) PAI as an inhibitor of MAO-B in the brain. Therefore, it is not effective in inhibiting the breakdown of brain dopamine, but is more active than the (R) isomer that causes intestinal smooth muscle relaxation. Therefore, [S] (-) PAI is not an inactive substance as found in racemic PAI when ingested with the (R) isomer.

Table 19: Antagonism of Bethanechol-induced contraction of the guinea pig ileal preparation by each of the two enantiomers of PAIHCl. A fixed dose of Bethanechol (0.8 µM) was administered in physiological solutions of the organ bath. A persistent contraction of the ileal stretch of a murine mounted mouse. Increasing addition of each of the two enantiomers of PAIHCl causes a dose-dependent relaxation of muscle. Results were expressed as percent relaxation relative to baseline (this was taken as 100% relaxation) before addition of histamine.

[0233]

[Table 19] Results [S] (−) PAI is inactive with [R] (+) PAI as an inhibitor of MAO-B in the brain. Therefore, it is not effective in inhibiting the breakdown of brain dopamine, but is more active than the (R) isomer that causes intestinal smooth muscle relaxation. Therefore, [S] (-) PAI is not an inactive substance as found in racemic PAI when ingested with the (R) isomer.

[Brief description of drawings]

FIG. 1 Example 22 showing MAO-A inhibitory activity in vitro
Is a graphical representation of the results according to.

FIG. 2 Example 22 showing MAO-B inhibitory activity in vitro
Is a graphical representation of the results according to.

FIG. 3A: Example 2 showing MAO activity in human cortical tissue
3 is a graph showing the results according to 2. Substrate is ¹⁴
Phenylethylamine (PEA) labeled with C.

FIG. 3B: Example 2 showing MAO activity in human cortical tissue
3 is a graph showing the results according to 2. Substrate is ¹⁴
5-hydroxytryptamine (5-H labeled with C
T).

FIG. 4. Short-term inhibition of MAO-A in the brain (ip).
Is a graphical representation of the results according to Example 23 showing.

FIG. 5. Short-term inhibition of MAO-B in the brain (ip).
Is a graphical representation of the results according to Example 23 showing.

FIG. 6: Short-term inhibition of MAO-A in the liver (i.
p. ) Is a graphical representation of the results according to Example 23.

FIG. 7. Short-term inhibition of MAO-B in the liver (i.
p. ) Is a graphical representation of the results according to Example 23.

FIG. 8 is a graphic representation of the results according to Example 23 showing the short term inhibition (per os) of MAO-A in the brain.

FIG. 9 is a graphical representation of results according to Example 23 showing short term inhibition (per os) of MAO-B in the brain.

FIG. 10. Short-term inhibition of MAO-A in the liver (per o
Figure 24 is a graphical representation of the results according to Example 23 showing s).

FIG. 11. Short-term inhibition of MAO-B in the liver (per o
Figure 24 is a graphical representation of the results according to Example 23 showing s).

FIG. 12: Long-term inhibition of MAO-A in the brain (per os)
25 is a graphical representation of the results according to Example 24 showing

FIG. 13: Long-term inhibition of MAO-B in the brain (per os)
25 is a graphical representation of the results according to Example 24 showing

FIG. 14: Long-term inhibition of MAO-A in the liver (per o
25 is a graphical representation of the results according to Example 24 showing s).

FIG. 15: Long-term inhibition of MAO-B in the liver (per o
25 is a graphical representation of the results according to Example 24 showing s).

FIG. 16 shows the i.d. of [R] (+) PAI. p. Figure 26 is a graphical representation of the results according to Example 25 showing MAO-B activity in rat brain as a function of time after administration.

FIG. 17 is a graph showing the results according to Example 32 showing the recovery of normal motion sickness (normokinesia) in mice administered with haloperidol 6 mg / kg sc. Mice received each test drug i.p. p. Was administered at the dose indicated by. Two hours later, they were administered haloperidol.
A kinetic score was scored 3 hours after haloperidol.
These scores consist of the ability to move horizontally along a rod, the ability to descend a vertical rod, and the shortening of catalepsy. In the absence of haloperidol, the maximum score is 12 and haloperidol alone is 6.8 ± 0.03. Statistical dominance was calculated by Student's "t" test. These are * p ≦ 0.05 versus haloperidol alone;
*** p ≦ 0.01; *** p ≦ 0.001.
[R] (+) PAI score is 5 mg / kg (p ≦ 0.0
5) 10 mg / kg (p ≦ 0.01) and 15 mg / kg (p
≦ 0.05) (n = 5.6), which is significantly different from the racemic PAI score. The doses given are for the free base of PAI (not for the mesylate salt).

FIG. 18: 100 mg / kg i. p. At α-methyl-
33 is a graphic representation of the results according to Example 32 showing the recovery of locomotor activity in rats treated with p-tyrosine. Rats received i.p. at the indicated doses. p. Administered the test compound. Two hours later, they were administered α-Mpt and quickly placed in the active cage. All locomotor activity was recorded for 10 hours. Control rats treated with saline recorded only 15,862 + 1424. With α-Mpt alone, the rat is 8,108 ± 810.
Was recorded. Statistical superiority according to Student's “t” test is * p ≦ 0.05; αp-Mpt; ** p ≦
0.01; *** p ≦ 0.001. [R] (+)
The PAI score is significantly different from the racemic PAI at 2 mg / kg (p ≦ 0.01), (n = 6). The doses given are for the free base of PAI,
Not a mesylate salt.

FIG. 19: NADH for 2 minutes of anoxia measured 30 minutes after injury and 30 minutes thereafter.
It is a graph which shows a response.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 25/28 A61P 25/28 25/30 25/30 25/32 25/32 27/02 27/02 43 / 00 111 43/00 111 (71) Applicant 591002197 TECHNION RESEARCH AND DEVELOPMENT FOUNDATION LIMITED TECHNION RESEARCH A NDDEVELOPMENT FOUND AT ION LIMITED Haifa 32000, TECHNION SITE, No Inventor 72, Senate House Mausa B.H.Yudim Haifa 32763, Hankin Street, Israel 18 (72) Inventor John P.M.Finberg Tibon, Israel 36 00, Ben Zubi Street 15 (72) Inventor Ruth Levy, Tel-Aviv 69415, Israel, Alterman Street 7 (72) Inventor Jeffrey Sterling, Israel, Jerusalem 97275, Ramot 156/1 ( 72) Inventor David Lerner Israel, Jerusalem 97241, Yegal Street 27/4 (72) Inventor Tartua Burger-Paskin Israel, Lana 43262, Akiba Street 16 (72) Inventor Heim Yerin Israel , Ramat-Gun 52333, Higherden Street 45 (72) Inventor Alex Beinberg, Rehoboth 76281, Israel, Ahaloni Street 5F Term (Reference) 4C206 AA01 AA02 FA07 MA01 MA04 NA14 ZA01 ZA02 ZA15 ZA16 ZA18 ZA33 ZC20 ZC39

Claims (2)

[Claims] 1. R (+)-N-propargyl-1-aminoindan or a pharmaceutically acceptable salt thereof is treated with cerebral ischemia, traumatic head injury, traumatic spinal cord injury, schizophrenia. , A pharmaceutical composition comprising as an active compound for the treatment of attention deficit, multiple sclerosis, withdrawal from addictive substances, or structural damage to the optic nerve. 2. The pharmaceutical composition according to claim 1, wherein the addictive substance is cocaine or alcohol.